Isolation of a chondroitin sulfate proteoglycan from a rat yolk sac tumor and immunochemical demonstration of its cell surface localization.
A proteoglycan was isolated from ascites fluid produced by a rat yolk sac tumor. The glycosaminoglycan chains of the proteoglycan are all sensitive to digestion with chondroitinase ABC and about 90% are sensitive to chondroitinase AC. The proteoglycan contains 5% protein. Amino acid analysis revealed a high content of serine and glycine which together constitute 37% of the amino acids. Glutamic acid (glutamine) and aspartic acid (asparagine) are also abundant. Galactosamine accounts for 97% of the hexosamine and the remainder is glucosamine. These characteristics indicate that the glycosaminoglycan side chains of this proteoglycan are predominantly chondroitin sulfate with a smaller amount of dermatan sulfate. Antibodies to the proteoglycan were prepared by immunization of a rabbit with purified alkali-treated proteoglycan. Affinity-purified antibodies from the antiserum immunoprecipitated (35S)sulfate-labeled radioactivity from culture media of the yolk sac tumor cells known to contain chondroitin sulfate proteoglycan. This binding was inhibited by the intact purified proteoglycan but not by proteoglycan treated with papain, suggesting dependence of the reactivity of the antibodies on integrity of the protein part of the proteoglycan. Immunofluorescence of the cultured yolk sac tumor cells revealed localization of immune reactive proteoglycans at the cell surface.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002807:Chondroitin / Q000031:analogs & derivatives*; D002809:Chondroitin Sulfates / Q000302:isolation & purification*; D005260:Female; D006025:Glycosaminoglycans / Q000302:isolation & purification*; D006595:Hexosamines / Q000032:analysis; D008649:Mesonephroma / Q000032:analysis*; D010051:Ovarian Neoplasms / Q000032:analysis*; D011509:Proteoglycans / Q000302:isolation & purification*; D011863:Radioimmunoassay; D051381:Rats; D014574:Uronic Acids / Q000032:analysis
D000596:Amino Acids; D006025:Glycosaminoglycans; D006595:Hexosamines; D011509:Proteoglycans; D014574:Uronic Acids; D002807:Chondroitin; D002809:Chondroitin Sulfates